title: Antithrombin attenuates vascular leakage via inhibiting neutrophil activation in acute lung injury.
abstract: OBJECTIVE
  To test the hypothesis that restoration of antithrombin plasma concentrations attenuates vascular leakage by inhibiting neutrophil activation through syndecan-4 receptor inhibition in an established ovine model of acute lung injury.
  
  
  METHODS
  Randomized controlled laboratory experiment.
  
  
  METHODS
  University animal research facility.
  
  
  METHODS
  Eighteen chronically instrumented sheep.
  
  
  METHODS
  Following combined burn and smoke inhalation injury (40% of total body surface area, third-degree flame burn; 4 × 12 breaths of cold cotton smoke), chronically instrumented sheep were randomly assigned to receive an IV infusion of 6 IU/kg/hr recombinant human antithrombin III or normal saline (n = 6 each) during the 48-hour study period. In addition, six sham animals (not injured, continuous infusion of vehicle) were used to obtain reference values for histological and immunohistochemical analyses.
  
  
  RESULTS
  Compared to control animals, recombinant human antithrombin III reduced the number of neutrophils per hour in the pulmonary lymph (p < 0.01 at 24 and 48 hr), alveolar neutrophil infiltration (p = 0.04), and pulmonary myeloperoxidase activity (p = 0.026). Flow cytometric analysis revealed a significant reduction of syndecan-4-positive neutrophils (p = 0.002 vs control at 24 hr). Treatment with recombinant human antithrombin III resulted in a reduction of pulmonary nitrosative stress (p = 0.002), airway obstruction (bronchi: p = 0.001, bronchioli: p = 0.013), parenchymal edema (p = 0.044), and lung bloodless wet-to-dry-weight ratio (p = 0.015). Clinically, recombinant human antithrombin III attenuated the increased pulmonary transvascular fluid flux (12-48 hr: p ≤ 0.001 vs control each) and the deteriorated pulmonary gas exchange (12-48 hr: p < 0.05 vs control each) without increasing the risk of bleeding.
  
  
  CONCLUSIONS
  The present study provides evidence for the interaction between antithrombin and neutrophils in vivo, its pathophysiological role in vascular leakage, and the therapeutic potential of recombinant human antithrombin III in a large animal model of acute lung injury.
authors.affiliation: 1Investigational Intensive Care Unit, Department of Anesthesiology, The University of Texas Medical Branch, Galveston, TX. 2Department of Anesthesiology, Intensive Care and Pain Medicine, University of Muenster, Muenster, Germany. 3Department of Pathology, The University of Texas Medical Branch, Galveston, TX. 4Shriners Hospital for Children, Galveston, TX.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Rehberg
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Yamamoto
authors.forename: Linda E
authors.initials: LE
authors.lastname: Sousse
authors.forename: Collette
authors.initials: C
authors.lastname: Jonkam
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhu
authors.forename: Lillian D
authors.initials: LD
authors.lastname: Traber
authors.forename: Robert A
authors.initials: RA
authors.lastname: Cox
authors.forename: Donald S
authors.initials: DS
authors.lastname: Prough
authors.forename: Daniel L
authors.initials: DL
authors.lastname: Traber
authors.forename: Perenlei
authors.initials: P
authors.lastname: Enkhbaatar
mesh_terms: D055371:Acute Lung Injury / Q000188:drug therapy* / Q000209:etiology / Q000473:pathology / Q000503:physiopathology*; D000402:Airway Obstruction / Q000188:drug therapy; D000818:Animals; D000990:Antithrombin III / Q000627:therapeutic use*; D000991:Antithrombins / Q000627:therapeutic use*; D002056:Burns / Q000150:complications; D002199:Capillary Permeability / Q000187:drug effects*; D002465:Cell Movement / Q000187:drug effects; D004195:Disease Models, Animal; D004487:Edema / Q000188:drug therapy; D005260:Female; D008168:Lung / Q000201:enzymology / Q000473:pathology; D018375:Neutrophil Activation / Q000187:drug effects*; D009504:Neutrophils / Q000378:metabolism / Q000502:physiology*; D009195:Peroxidase / Q000378:metabolism; D011659:Pulmonary Gas Exchange / Q000187:drug effects; D011897:Random Allocation; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism*; D011994:Recombinant Proteins / Q000627:therapeutic use; D012756:Sheep; D015208:Smoke Inhalation Injury / Q000150:complications; D053671:Syndecan-4 / Q000378:metabolism
chemical_list: D000991:Antithrombins; D043562:Receptors, G-Protein-Coupled; D011994:Recombinant Proteins; D053671:Syndecan-4; D000990:Antithrombin III; D009195:Peroxidase